Significance of indoor Significance of indoor air PM air PM
exposureexposure
•• Indoor environments: variable and potentially Indoor environments: variable and potentially distinct sources of chemical exposuredistinct sources of chemical exposure
•• High concentrationsHigh concentrations
•• Close to the sourceClose to the source
•• Time of exposure Time of exposure
•• Small databaseSmall database
Sources ofSources of
PM from Indoor AirPM from Indoor Air
•• Wood burningWood burning
•• CandlesCandles
•• IncenseIncense
•• Cooking activities: Frying, Broiling, Oven etc.Cooking activities: Frying, Broiling, Oven etc.
•• FlooringFlooring
IndoorIndoor PM exposurePM exposurePossible Health EffectsPossible Health Effects
•• Airway Symptoms and AllergyAirway Symptoms and Allergy
•• Cardiovascular diseases (CVD) like Cardiovascular diseases (CVD) like atherosclerosis atherosclerosis
•• Endocrine disrupting effectsEndocrine disrupting effects
•• Immunotoxic effectsImmunotoxic effects
•• Chronic inflammatory response as a main Chronic inflammatory response as a main cause for adverse health effectscause for adverse health effects
In vitro Cell ModelIn vitro Cell Model•• Critical sensors for smoke, stress, and PM Critical sensors for smoke, stress, and PM
exposure exposure •• ArylhydrocarbonArylhydrocarbon--Receptor (AhR): LigandReceptor (AhR): Ligand--
activated through numerous planar aromatic activated through numerous planar aromatic hydrocarbons as well as stresshydrocarbons as well as stress
•• TollToll--like receptors (TLR): Pathogen like receptors (TLR): Pathogen recognition and activation of innate immunity recognition and activation of innate immunity
•• Nuclear factorNuclear factor--kappa B (NFkappa B (NF--ĸĸB): Sensor for B): Sensor for smoke and stresssmoke and stress
Study ObjectiveStudy ObjectiveSource:
•cooking
•candles
•wood smoke
•incense
Filter
ChemistryBioassay
NCI H441U937
Real-time PAH
•screeningVapor Collection
Selective Ion Monitoring (SIM)
•quantitativeCYP1A1
Total Ion Chromatogram (TIC)
•qualitativeIL-8COX-2 CYP1A1
COX-2MUC5AC
Add internal standard
Gene expression
Extraction Extraction
Gene expression
Source:
•cooking
•candles
•wood smoke
•incense
Filter
ChemistryBioassay
NCI H441U937
Real-time PAH
•screeningVapor Collection
Selective Ion Monitoring (SIM)
•quantitativeCYP1A1
Total Ion Chromatogram (TIC)
•qualitativeIL-8COX-2 CYP1A1
COX-2MUC5AC
Add internal standard
Gene expression
Extraction Extraction
Gene expression
Macrophages Lung Cells
Study DesignStudy Design
•• Two main target cell types Two main target cell types
a) Phagocytotic cell types as first line of defense, a) Phagocytotic cell types as first line of defense,
human macrophages (U937) human macrophages (U937)
b) Lung Clara cells from pulmonary epithelium (NCI b) Lung Clara cells from pulmonary epithelium (NCI
H441) H441)
Biomarkers of PM exposureBiomarkers of PM exposure
•• CYP1A1: Cytochrome P450 monooxygenase, xenobiotic CYP1A1: Cytochrome P450 monooxygenase, xenobiotic
metabolizing enzyme, bioactivation, Ahmetabolizing enzyme, bioactivation, Ah--Receptor Receptor
regulatedregulated
•• COXCOX--2: Cyclooxygenase, key enzyme for production of 2: Cyclooxygenase, key enzyme for production of
prostaglandins involved in inflammation, upregulated in prostaglandins involved in inflammation, upregulated in
cancer cellscancer cells
•• ILIL--8: Interleukin 8, chemoattractant peptide for 8: Interleukin 8, chemoattractant peptide for
neutrophils, major mediator of inflammatory responseneutrophils, major mediator of inflammatory response
•• HOHO--1: Hemeoxygenase 1, essential enzyme in heme 1: Hemeoxygenase 1, essential enzyme in heme
catabolism, protect cells against oxidative injury. HOcatabolism, protect cells against oxidative injury. HO--1 is 1 is
a stressa stress--responsive protein and induced by exposure toresponsive protein and induced by exposure to
various forms of oxidative stressvarious forms of oxidative stress
COX-2CYP1A1
IL-8HO-1
PMs
Macrophage ModelMacrophage Model
PMs containing
PAHs, Dioxins,PCBs,Metals,
Endotoxin
Macrophage cell
Release of Cytokines Activated inflammatory
Macrophage
Accumulation of CholesterolFoam cell formation
AhR
TLR
NF-κB
Macrophage ModelMacrophage Model
•• Macrophage as a sensor of danger in the hostMacrophage as a sensor of danger in the host
•• MaMacrophages have remarkable plasticity and crophages have remarkable plasticity and change their physiology in response to change their physiology in response to environmental stimulienvironmental stimuli
•• Foam cell formation as precursor of CVD Foam cell formation as precursor of CVD
Macrophage modelMacrophage model--U937 MonocyteU937 Monocyte
U937 macrophagesU937 macrophages
Activated U937 macrophages (Foam cells)Activated U937 macrophages (Foam cells)
Formation of foam cellsFormation of foam cells
Swelling of the intima in the wall of the artery which pushes the endothelium into the lumen of the artery
Arterial wall
Foam cells
Control U937monocyte Control MØ Dioxin MØ
Development of atherosclerotic lesions in Development of atherosclerotic lesions in
ApoE miceApoE mice
ApoE, Ctrl ApoE, Dioxin (TCDD)
Filtered Air PM 2.5
Lung Clara Cell ModelLung Clara Cell Model
•• NCI H441 Clara cell characteristicsNCI H441 Clara cell characteristics
•• Primary secretory cells in small airwaysPrimary secretory cells in small airways
•• Clara cells can differentiate into mucin Clara cells can differentiate into mucin producing Goblet cellsproducing Goblet cells
•• Pathological feature of asthma and COPD Pathological feature of asthma and COPD for adverse health effectsfor adverse health effects
NCI H441 Lung Clara cell modelNCI H441 Lung Clara cell model
PM/PAH
PM uptake MUC5AC- Mucin-induction accumulation
• Chronic obstructive pulmonary disease (COPD)• Emphysema• Asthma
COXCOX--2 Response of various cell lines 2 Response of various cell lines
MUC5AC Response of various cell lines MUC5AC Response of various cell lines
CookingCooking
CookingCooking
Cooking PM SamplingCooking PM Sampling
Stir-fry Oven
Effect of cooking PM in Macrophages Effect of cooking PM in Macrophages
C: Vehicle control
FBL-2: Field Filter Blank
BGA: Background Air PM10 filter
CK-1:Stir-fry cooking
CK-2: Oven cooking
Effect of cooking PM in Lung CellsEffect of cooking PM in Lung Cells
C: Vehicle control
FBL-2: Field Filter Blank
BGA: Background Air PM10 filter
CK-1:Stir-fry cooking
CK-2: Oven cooking
CandleCandle
Variety of CandlesVariety of Candles
Candle ID Name Characteristics Color/Style Size Store Manufactured In
A Tindra Ljuv Scented Candle Red, Filled Glass
2.5" dia X 1.8" 2 China
B 3'' Fresh Cotton Scented Candle White, Pillar2 3/4'' (D) X
3''(H)3 India
CBotanica Candles
Scented and Handcrafted Candle, Mango Papaya
Orange Red, Pillar
260 g, 2 7/8'' (D) X 3''
(H)4 Hong Kong
D Paula DeenScented Candle, Pear Honey
Green, Filled Glass (Container)
16 oz (453 g) 1 USA
EScented Gold Ring Pink
Scented Gold Ring, Religious
Flamingo, Filled glass
2 1/4'' (D) x 8 '' (H)
2 USA
F RenewHand poured, Jasmine & Tea Leaf
Coral, Pillar8.8 oz/250 g,
2.75'' x 3 ''4 Vietnam
Effect of candle PM in MacrophagesEffect of candle PM in Macrophages
Bkgrd: Background Air
CN-A to CN-F: Candle samples
Effect of candle PM in Lung cellsEffect of candle PM in Lung cells
Bkgrd: Background Air
CN-A to CN-F: Candle samples
Effect of candle PM RetestEffect of candle PM Retest
C: Vehicle Control
BGA: Background Air
CN-A to CN-B: Candle samples
Wood SmokeWood Smoke
Wood Smoke PM SamplingWood Smoke PM Sampling
Fireplace
Cyclone
XAD
Filter Pak
Vacuum Pump
Effect of Wood smoke PM on CYP1A1 Effect of Wood smoke PM on CYP1A1
in Macrophagesin Macrophages
FBL: Filter Blank, BA ws: Background Air PM 2.5, BA10ws: Background Air PM10WS-1: Woodsmoke PM2.5 run#1, WS10-1: Woodsmoke PM10 run # 1WS-2: Woodsmoke PM2.5 run#2, WS10-2: Woodsmoke PM10 run # 2UDP: Urban Dust NIST SRM 1649, DEP: Diesel PM NIST SRM2975
Effect of Wood smoke PM on ILEffect of Wood smoke PM on IL--8 in 8 in
MacrophagesMacrophages
FBL: Filter Blank, BA ws: Background Air PM 2.5, BA10ws: Background Air PM10WS-1: Woodsmoke PM2.5 run#1, WS10-1: Woodsmoke PM10 run # 1WS-2: Woodsmoke PM2.5 run#2, WS10-2: Woodsmoke PM10 run # 2UDP: Urban Dust NIST SRM 1649, DEP: Diesel PM NIST SRM2975
Effect of Wood smoke PM on CYP1A1 Effect of Wood smoke PM on CYP1A1
in Lung Cellsin Lung Cells
FBL: Filter Blank, BA ws: Background Air PM 2.5, BA10ws: Background Air PM10WS-1: Woodsmoke PM2.5 run#1, WS10-1: Woodsmoke PM10 run # 1WS-2: Woodsmoke PM2.5 run#2, WS10-2: Woodsmoke PM10 run # 2UDP: Urban Dust NIST SRM 1649, DEP: Diesel PM NIST SRM2975
Effect of Wood smoke PM on MUC5AC Effect of Wood smoke PM on MUC5AC
in Lung Cellsin Lung Cells
FBL: Filter Blank, BA ws: Background Air PM 2.5, BA10ws: Background Air PM10WS-1: Woodsmoke PM2.5 run#1, WS10-1: Woodsmoke PM10 run # 1WS-2: Woodsmoke PM2.5 run#2, WS10-2: Woodsmoke PM10 run # 2UDP: Urban Dust NIST SRM 1649, DEP: Diesel PM NIST SRM2975
IncenseIncense
Variety of IncensesVariety of Incenses
Sample ID Name Color Size Type Store Manufactured
A Nag Champa Brown 15g Net weight Wood core 1Bangalore, India
BPure Tibetan-Herbal Medicine
Brown N/A No Core 1Kathmandu, Nepal
CShoyeido Traditional Japanese
Multi.017 oz per stick, 10 sticks/Pack
No Core 1 Kyoto, Japan
DPure Tibetan-Potala
Red N/A No Core 1 Nepal
EAromatherapy variety
Multi10 in, 24 sticks/Pack
Wood core 2 Mumbai, India
FJoss Sticks Mainichikoh
Green 107 sticks No Core 3 Japan
G Floral variety Multi10 in, 24 sticks/Pack
Wood core 2 Mumbai, India
Incense PM SamplingIncense PM Sampling
Cyclone
Vacuum Pump
70 mm Filter Pak
Fume Hood Incense
Effect of Incense PM in MacrophagesEffect of Incense PM in Macrophages
C: DMSO ControlBGA: Background AirINC-A: Incense Nag ChampaINC-B: Incense TibetINC-C: Incense ShoyiedoINC-D: Incense PotalaINC-E: Incense AromatherapyINC-F: Joss Stick greenINC-G: Floral Variety
DoseDose--response of Incense PM in Macrophagesresponse of Incense PM in Macrophages
C: DMSO Control, PBL: Process Blank, INC-Fa: Joss Stick green PM10INC-Fb: Joss Stick green PM2.5, INC-Gb: Floral Variety PM2.5
DoseDose--response of Incense PM in Lung Cellsresponse of Incense PM in Lung Cells
C: DMSO Control, PBL: Process Blank, INC-Fa: Joss Stick green PM10INC-Fb: Joss Stick green PM2.5, INC-Gb: Floral Variety PM2.5
Comparison of PM sample groups Comparison of PM sample groups
PAHs in Incense PM FractionsPAHs in Incense PM Fractions
Fraction 1 Fraction 4Fraction 3Fraction 2
Chemical fractions F1 to F4: non-polar > polar
Effect of incense PM fractions on Effect of incense PM fractions on
CYP1A1CYP1A1
Chemical fractions F1 to F4: non-polar > polar; neat: un-fractionatedINC-Fa: PM2.5 Incense, Mainichi-koh Japanese-1, INC-Ga: PM2.5 Incense,
Floral India-1, INC-Gb: PM2.5 Incense, Floral India-2 duplicateDEP: Diesel engine exhaust particles (NIST 2975)
Effect of incense PM fractions on COXEffect of incense PM fractions on COX--
22
Chemical fractions F1 to F4: non-polar > polar; neat: un-fractionatedINC-Fa: PM2.5 Incense, Mainichi-koh Japanese-1, INC-Ga: PM2.5
Incense, Floral India-1, INC-Gb: PM2.5 Incense, Floral India-2 duplicateDEP: Diesel engine exhaust particles (NIST 2975)
Summary 1Summary 1•• The human Macrophage Cell line The human Macrophage Cell line
(U937)and Lung Clara cell (NCIH441) (U937)and Lung Clara cell (NCIH441) suitable in vitro model for PM testingsuitable in vitro model for PM testing
•• All indoor PMs tested generated a All indoor PMs tested generated a response in at least one or both cell typesresponse in at least one or both cell types
•• Wood Smoke samples increase CYP1A1 Wood Smoke samples increase CYP1A1 and COXand COX--2 significantly in Macrophages 2 significantly in Macrophages
•• All incenses were tested very active in the All incenses were tested very active in the induction of biomarkers (polar fractions induction of biomarkers (polar fractions more active)more active)
Summary 2Summary 2
•• Effective and inexpensive indoor air Effective and inexpensive indoor air monitoringmonitoring
•• Assessment of mixture toxicity Assessment of mixture toxicity (combined effect of exposure)(combined effect of exposure)
•• Assessment of exposure and Assessment of exposure and evaluation of health effectsevaluation of health effects
Cleaner Indoor AirCleaner Indoor Air
Thank youThank you
Lori Miyasato
Peter Mathews
Norm Kado
Reiko Kobayashi
Xiaoxue Liu
Rupinder Kaur
Teresa Chiang
Noriko Nishimura
Patrick Wong
Dalei Wu
Wen Li
Viktoria Kuo
Patty Lok
Danitza Alvizar
Helen Woldai
Fumio Matsumura